• Asian Pacific Journal of Cancer Prevention
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• 제14권4호
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• pp.2311-2315
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• 2013

Objective: To investigate the expression level of TUG1 and one of its transcript variants (n377360) in osteosarcoma cells and assess the role of TUG1 in proliferation and apoptosis in the U2OS cell line. Methods: TUG1 and n377360 expression levels in patients with osteosarcomas and the U2OS human osteosarcoma cell line were evaluated using real-time quantitative PCR. U2OS cells were transected with TUG1 and n377360 siRNA or non-targeting siRNA. MTS was performed to assess the cell proliferation and flow cytometry was applied to analyze apoptosis. Results: We found significantly higher TUG1 and n377360 expression levels in osteosarcoma tissues compared with matched non-tumorous tissues. In line with this, suppression of TUG1 and n377360 expression by siRNA significantly impaired the cell proliferation potential of osteosarcoma cells. Furthermore, inhibition of TUG1 expression significantly promoted osteosarcoma cell apoptosis. Conclusions: The overexpression of TUG1 and n377360 in osteosarcoma specimens and the functional role of TUG1 and n377360 regarding cell proliferation and apoptosis in an osteosarcoma cell line provided evidence that the use of TUG1 or n377360 may be a viable but an as yet unexplored therapeutic strategy in tumors that over express these factors.

• Asian Pacific Journal of Cancer Prevention
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• 제14권6호
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• pp.3751-3755
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• 2013

Backgroud and Aims: MicroRNA-206 has proven to be down-regulated in many human malignancies in correlation with tumour progression. Our study aimed to characterize miR-206 contributions to initiation and malignant progression of human osteosarcoma. Methods: MiR-206 expression was detected in human osteosarcoma cell 1ine MG63, human normal osteoblastic cell line hFOB 1.19, and paired osteosarcoma and normal adjacent tissues from 65 patients using quantitative RT-PCR. Relationships of miR-206 levels to clinicopathological characteristics were also investigated. Moreover, miR-206 mimics and negative control siRNA were transfected into MG63 cells to observe effects on cell viability, apoptosis, invasion and migration. Results: We found that miR-206 was down-regulated in the osteosarcoma cell line MG63 and primary tumor samples, and decreased miR-206 expression was significantly associated with advanced clinical stage, T classification, metastasis and poor histological differentiation. Additionally, transfection of miR-206 mimics could reduce MG-63 cell viability, promote cell apoptosis, and inhibit cell invasion and migration. Conclusions: These findings indicate that miR-206 may have a key role in osteosarcoma pathogenesis and development. It could serve as a useful biomarker for prediction of osteosarcoma progression, and provide a potential target for gene therapy.

• Asian Pacific Journal of Cancer Prevention
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• 제14권8호
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• pp.4615-4619
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• 2013

Background: Artesunate, extracted from Artemisia annua, has been proven to have anti-cancer potential. Allicin, diallyl thiosulfinate, the main biologically active compound derived from garlic, is also of interest in cancer treatment research. This object of this report was to document synergistic effects of artesunate combined with allicin on osteosarcoma cell lines in vitro and in vivo. Methods: After treatment with artesunate and allicin at various concentrations, the viability of osteosarcoma cells was analyzed by MTT method, with assessment of invasion and motility, colony formation and apoptosis. Western Blotting was performed to determine the expression of caspase-3/9, and activity was also detected after drug treatment. Moreover, in a nude mouse model established with orthotopic xenograft tumors, tumor weight and volume were monitored after drug administration via the intraperitoneal (i.p.) route. Results: The viability of osteosarcoma cells in the combination group was significantly decreased in a concentration and time dependent manner; moreover, invasion, motility and colony formation ability were significantly suppressed and the apoptotic rate was significantly increased through caspase-3/9 expression and activity enhancement in the combination group. Furthermore, suppression of tumor growth was evident in vivo. Conclusion: Our results indicated that artesunate and allicin in combination exert synergistic effects on osteosarcoma cell proliferation and apoptosis.

• Asian Pacific Journal of Cancer Prevention
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• 제14권8호
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• pp.4651-4654
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• 2013

Choroquine (CQ) and valproic acid (VPA) have been extensively studied for biological effects. Here, we focused on efficacy of combined CQ and VPA on osteosarcoma cell lines. Viability of osteosarcoma cell lines (U20S and HOS) was analyzed by MTT assay. Apoptotic assays and colony formation assays were also applied. ROS generation and Western Blotting were performed to determine the mechanism of CQ and VPA combination in the process of apoptosis. The viability of different osteosarcoma cell lines significantly decreased after CQ and VPA combination treatment compared with either drug used alone, and apoptosis was increased significantly. ROS generation was triggered leading to expression of apoptosis related genes being increased and of antiapoptotic related genes being decreased. From our data shown here, CQ and VPA combination treatment in vitro enhanced cytotoxicy to osteosarcoma cells.

• 대한골관절종양학회지
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• 제13권2호
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• pp.88-95
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• 2007

목적: wild-type p53 단백질을 가지고 있는 U2OS 인간 골육종 세포주의 세포자멸사에서 MMP억제제와 doxorubicin의 작용을 비교하고자 하였다. 대상 및 방법: 배양한U2OS 세포주에 Doxorubicin, MMP억제제(MMPI III)를 따로 투여한 것과 두 약물을 동시에 투여한 것을 비교하였다. 또한 Doxorubicin의 작용이 Fas/FasL 경로를 통해 유발되는지 알아보기 위해 Fas 중화항체를 doxorubicin과 함께 투여하였다. 약물을 처리하여 배양한 세포에서 발생한 세포자멸사와 세포괴사를 확인하기 위해서 유세포 검사를 시행하였다. 결과: Doxorubicin을 처리한 세포가 처리하지 않은 세포보다 약물 농도에서 더 많은 세포괴사(p=0.000)를 보였다. 이에 반해 MMPI III를 처리한 세포는 약물 농도와 반응시간에 따른 세포자멸사 및 세포괴사에서 차이를 보이지 않았다. 두 약물을 동시에 투여한 군은 그렇지 않은 군에 비해 세포 자멸사 및 세포괴사에서 차이가 없었다. Doxorubicin에 Fas 중화항체를 추가한 군과 doxorubicin 단독 투여 군 사이에 세포자멸사 및 세포괴사에서 차이가 없었다. 결론: Doxorubicin과 같이 MMP 억제제를 골육종 치료에 사용하려면 wild-type p53 뿐만 아니라 wild-type p14를 가지고 있는 골육종 세포에 대한 검사가 필요할 것으로 사료된다.

• Molecules and Cells
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• 제38권7호
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• pp.638-642
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• 2015

Quercetin can suppress osteosarcoma cell growth and metastasis. However, other effects of quercetin on osteosarcoma remain largely unknown. This research aims to evaluate the effects of quercetin in combination with cisplatin as treatment for osteosarcoma and investigate its regulatory mechanism. Cell viability and apoptosis in 143B cell line were determined after treatment with quercetin and/or cisplatin. RT-PCR and Western blot analysis were performed to determine the RNA or protein expression levels. Moreover, transwell assay was used to evaluate metastasis. Furthermore, rescue experiments were performed to investigate the potential regulatory mechanism of the treatment. Results showed that quercetin with concentration that was equal to or greater than $10{\mu}M$ inhibited 143B proliferation, while $5{\mu}M$ quercetin enhanced the cisplatin sensitivity of 143B cells. Expression of miR-217 was upregulated after quercetin and/or cisplatin treatment, while its target KRAS was downregulated both at mRNA and protein levels. MiR-217 knockdown led to the loss of enhanced cisplatin sensitivity while miR-217 overexpression showed the opposite effects, indicating that quercetin regulated cisplatin sensitivity by modulating the miR-217-KRAS axis. In conclusion, $5{\mu}M$ quercetin enhanced the cisplatin sensitivity by modulating the miR-217-KRAS axis. This finding suggests that quercetin may be administered with cisplatin to improve the treatment for osteosarcoma.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.795-799
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• 2013

Ku70/80 heterodimer is a central element in the nonhomologous end joining (NHEJ) DNA repair pathway, Ku80 playing a key role in regulating the multiple functions of Ku proteins. It has been found that the Ku80 protein located at telomeres is a major contributor to radiosensitivity in some telomerase positive human cancer cells. However, in ALT human osteosarcoma cells, the precise function in radiosensitivity and telomere maintenance is still unknown. The aim of this study was to investigate the effects of Ku80 depletion in the U2OS ALT cell line cell line. Suppression of Ku80 expression was performed using a vector-based shRNA and stable Ku80 knockdown in cells was verified by Western blotting. U2OS cells treated with shRNA-Ku80 showed lower radiobiological parameters (D0, Dq and SF2) in clonogenic assays. Furthermore, shRNA-Ku80 vector transfected cells displayed shortening of the telomere length and showed less expression of TRF2 protein. These results demonstrated that down-regulation of Ku80 can sensitize ALT cells U2OS to radiation, and this radiosensitization is related to telomere length shortening.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10797-10801
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• 2015

Background: To explore the molecular mechanisms of metastatic osteosarcoma (OS) by using the microarray expression profiles of metastatic and non-metastatic OS samples. Materials and Methods: The gene expression profile GSE37552 was downloaded from Gene Expression Omnibus database, including 2 human metastatic OS cell line models and 2 two non-metastatic OS cell line models. The differentially expressed genes (DEGs) were identified by Multtest package in R language. In addition, functional enrichment analysis of the DEGs was performed by WebGestalt, and the protein-protein interaction (PPI) networks were constructed by Hitpredict, then the signal pathways of the genes involved in the networks were performed by Kyoto Encyclopaedia of Genes and Genomes (KEGG) automatic annotation server (KAAS). Results: A total of 237 genes were classified as DEGs in metastatic OS. The most significant up- and down-regulated genes were A2M (alpha-2-macroglobulin) and BCAN (brevican). The DEGs were significantly related to the response to hormone stimulus, and the PPI network of A2M contained IL1B (interleukin), LRP1 (low-density lipoprotein receptor-related protein 1) and PDGF (platelet-derived growth factor). Furthermore, the MAPK signaling pathway and focal adhesion were significantly enriched. Conclusions: A2M and its interactive proteins, such as IL1B, LRP1 and PDGF may be candidate target molecules to monitor, diagnose and treat metastatic OS. The response to hormone stimulus, MAPK signaling pathway and focal adhesion may play important roles in metastatic OS.

• 대한골관절종양학회지
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• 제15권2호
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• pp.111-121
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• 2009

배경: 골육종은 소아 및 청소년기에 가장 흔하게 발생하는 악성 골종양 중 하나이다. 최근 수술적 치료와 항암 화학요법을 병행하여 생존률이 증가하였지만, 아직까지도 항암제는 항암제 내성 및 이차성 악성 종양의 발생 등 여러 문제점을 가지고 있다. 일부 종양은 matrix metalloproteinase(MMPs)의 발현이 증가되어 있고, MMP inhibitor에 대한 연구가 진행되고 있다. 반면 bisphosphonate(BPs) 제제는 골흡수를 억제하는 능력이 있으며, 파골세포와 관계된 골 병변에 광범위하게 사용되고 있다. 또 bisphosphonate 제제는 직접적인 항암효과를 가지고 있는 것으로 알려져 있다. 대상 및 방법: 골육종 세포주(U2OS)를 ibandronate(0, 0.1, 1, 10M)를 이용하여 48시간 동안 처치하였다. 세포의 생존능은 MTT assay를 이용하여, MMP-2와 MT1-MMP의 mRNA level은 reverse-transciption polymerase chain reaction을 이용하여 측정하였으며, MMP-2와 MT1-MMP 단백의 양은 Westernblot을, MMP-2의 활성은 Gelatin zymography를 이용하여 측정하였다. 또, ibandronate 처치 전후의 골육종 세포주의 침습성은 Matrigel invasion assay를 이용하여 측정하였다. 결과: 48시간 ibandronate 처치 후 U2OS 세포주의 침습력은 ibandronate에 대해 용량 의존적으로 감소하였다. 특히 10M 이내의 ibandronate는 세포독성이 나타나지 않았다. 젤라틴 융해능과 MMP-2 및 MT1-MMP의 단백 및 mRNA 정도역시 ibandronate 농도가 증가할수록 감소하였다. 결론: Ibandronate는 골육종 세포주의 MMP-2 및 MT1-MMP의 발현을 억제하였으며, 종양세포의 침습력을 감소시켰다. Bisphosphate의 종양세포 침습 억제 능력은 새로운 전이 억제제의 개발에 도움이 될 것으로 사료된다.

• 치과방사선
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• 제26권2호
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• pp.121-132
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• 1996

The purpose of this study was to aid in the prediction of tumor cell tolerance to radiotherapy and/or chemotherapy. For this study, cell surviving curves were obtained for human osteosarcoma MG-63 cell line using semiautomated MTT assay. 2,4, 6, 8, 10Gy were irradiated at a dose rate of 210cGy/min using /sup 60/Co Irradiator ALDORADO 8. After irradiation, MG-63 cell lines(3×10⁴ cells/ml) were exposed to bleomycin and cisplatin at concentration of 0.2, 2, 20㎍/㎖ for 1 hour respectively. The viable cells were determined for each radiation dose and/or each concentration of drug. And they were compared to control values. The obtained results were as follows : 1. There was significant difference of surviving fraction at 4, 6, 8, 10Gy on MG-63 cell line(p<0.05). 2. There was significant difference of cytotoxicity of bleomycin or cisplatin at all concentration of 0.2, 2, 20㎍/㎖ (p<0.05) on MG-63 cell line. The cytotoxicity of cisplatin was more effective than bleomycin at concentration of 20㎍/㎖ on MG-63 cell line. 3. There was significant difference of cytotoxicity of bleomycin or cisplatin at all concentration after irradiation of 2Gy on MG-63 cell line. 4. There was significant difference of cytotoxicity of bloemycin or cisplatin at concentration of 20㎍/㎖ after irradiation than that of irradiation alone(p<0.01). But there was no significant difference of cytotoxicity of bleomycin at concentration of 20㎍/㎖ after irradiation of l0Gy than that of irradiation alone.